Investigate Inhibitory Effects of Ginger polyphenols compare to Simvastatin towards HMG-CoA reductase: An Integrated Molecular Docking and Molecular dynamic simulation

Diabetes is an increasing problem in Ethiopia, affecting up to 6.5% of Ethiopian adults. There are serious complications associated with diabetes including macrovascular and microvascular. Controlling Lipid profiles and blood glucose significantly reduces the risk of complications. Statins are the only current treatment for both type 1 diabetes and Type 2 diabetes dyslipidemia. HMG-CoA reductase plays a central role in the production of cholesterol which, associated with cardiovascular disease (CVD). Statins have been found to reduce cardiovascular disease and mortality in those who are at high risk. Nonetheless, it has adverse effect, such as drug-related hypoglycemia and high cost. These situations lead to develop suitable phytotherapeutic agents with less frequent side effects. Ginger (Zingiber officinale) is widely consumed as a spice, and numerous studies suggest that ginger may have beneficial effects for diabetes and dyslipidemia. But, further studies are needed to investigate effects of binding affinity and binding site residues for major ginger extract polyphenols towards target HMG-CoA reductase. In this study, ADMET web server, Auto-Dock 5.4 and Gromacs 2020 were used. Out of eleven major gingers polyphenols screened three selected based on docking energy compare to Simvastatin for MD simulation. The predicted binding affinity for 6-paradol, 6-shogaol and gingerdione were −8.51, −6.93, −9.24 kcal/mol, respectively. The results of molecular dynamic simulation are consistence with docking. The predicted ligand binding site residues are Arg641, Gly808, Arg641, Met781, Ser794 and Arg595. In conclusion, 6-paradol, 6-shogaol and gingerdione could be possible therapy because, of interactions with target HMG-CoA reductase. Therefore, further wet lab study will be needed, for the better understanding of the mechanism of action of ginger extract by which it modulates liver and kidney vivo condition.

mortality in those who are at high risk. Nonetheless, it has adverse effect, such as drug-related 23 hypoglycemia and high cost. These situations lead to develop suitable phytotherapeutic agents 24 with less frequent side effects. Ginger (Zingiber officinale) is widely consumed as a spice, and 25 numerous studies suggest that ginger may have beneficial effects for diabetes and dyslipidemia. 26 But, further studies are needed to investigate effects of binding affinity and binding site residues 27 for major ginger extract polyphenols towards target HMG-CoA reductase. In this study, ADMET 28 web server, Auto-Dock 5.4 and Gromacs 2020 were used. Out of eleven major gingers 29 polyphenols screened three selected based on docking energy compare to Simvastatin for MD 30 simulation. The predicted binding affinity for 6-paradol, 6-shogaol and gingerdione were -8.51, -31 6.93, -9.24 kcal/mol, respectively. The results of molecular dynamic simulation are consistence 32 with docking. The predicted ligand binding site residues are Arg641, Gly808, Arg641, Met781, 33 Ser794 and Arg595. In conclusion, 6-paradol, 6-shogaol and gingerdione could be possible 34 therapy because, of interactions with Diabetes mellitus is the most common metabolic and endocrine disorder worldwide. It is linked 42 to disturbances in carbohydrate, fat and protein metabolism and important study area because of 43 the global prevalence of diabetes is projected to rise in coming years [1] . It is an increasing 44 problem that contributes significant mortality from increased microvascular and macrovascular And also, an estimated 700 million adults worldwide will have diabetes by 2045 [3 6] . Over the 50 last 30 years, type 2 diabetes has changed from relatively mild ailment associated with aging to 51 one of a major cause of premature mortality and morbidity in most countries [  AdmetSAR 2.0 webserver [38] and swissADMET webserver to computed blood brain barrier 119 (BBB) permeant, gastro-intestinal (GI) absorption, cytochrome inhibition. Finally, based on 120 ADME prediction, ligands were made ready for molecular docking analysis using Auto Dock  Ramachandran plot analysis by using UCLA-DOE server-web to assess the overall quality and 127 local quality analysis, based on the Z-score and knowledge-based energy calculations [39] .

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Validate the modeled structure using the UCLA-DOE server (http://servicesn.mbi.ucla.edu/) [18 19 129 20] . The modeled 3-D structure was then validated and confirmed by using the RAMPAGE, 130 ERRAT, and Verify 3-D online servers. Based on UCLA-DOE server-web results missed 131 residues and atoms were fixed by using Swiss-pdb viewer and rescreened Structural evaluation 132 and stereo chemical quality [39] . Finally, the .pdb file was entered into Auto Dock 4.2 for  Entire simulations are carried out using GAFF force field [42] . The System was placed at center of       properties and Lipinski's rule 5 prediction and the results are tabulated (Table 1 and Table 2).

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The pharmacokinetic properties and Lipinski's rule 5 of top-three compounds close to co-crystal 244 reference drug as shown in Table 1 and Table 2. In this study, reference drug consist of 7 245 torsional angles in its structure, while the three ginger extract consist of rotatable bonds in the 246 range of 9-14. The Hydrogen bond acceptor (HBA) and donor (HBD) profiles of three selected 247 ginger extract close to reference drug. ADMET for top-five compounds (Hexahydrocurcumin, 248 10-shogaol, 10-gingerol, 8-shogaol and 6-shogaol) close to reference drug as shown in Table 2.

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Simultaneously, before going for docking, three-dimensional crystal structure of the HMG-CoA

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(3-hydroxy-3-methylglutaryl-coenzyme A) reductase (PDB Id: 1hw9) was analyzed by using the residues Arg641, Arg641, Met781, Ser794 and Arg595, which indicates the importance of these residues to maintain HMG-CoA reductase-ginger extracts complex (Table 3). Except, all 289 the ginger extracts capable of forming hydrogen bonds with HMG-CoA reductase (Table 3 290 supportive). Overall molecular docking study showed that the compound simvastatin had highest 291 binding affinity that can produce the inhibition even at low concentration, hence, we have 292 prioritized simvastatin as the best candidate for further optimization for in-vitro studies. which is consistence with previous study [15] . Last but not least, the radius of gyration (Rg) value From these research reports one might predict that lowering the lipids profile would prevent the 321 progression of plague formation. However, to date, lipids profile-specific drug is not available 322 and the effect of routinely used cardiovascular drugs on HMG-CoA reductase is not known.

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Hence, it is very essential to study the association between known cardio-protective drugs such 324 as simvastatin and HMG-CoA reductase. To our knowledge, this is the first study which has 325 proposed the three-dimensional structure of HMG-CoA reductase with possible binding site 326 residues and predicted the binding efficiency of ginger extracts towards HMG-CoA reductase.